Laboratory diagnosis of brucellosis in a rural endemic area in northeastern Spain

Sera obtained from 62 patients from four mountain counties in Catalonia (Northeastern Spain), in whom brucellosis had been diagnosed on the basis of clinical evidence and/or personal history, were analyzed using the rose Bengal test, standard serum agglutination test (SAT), Coombs" test, ELISA, and complement fixation. The diagnosis was further confirmed through blood cultures. Clinical evidence, epidemiology, and the results from serologic tests were used to assign patients to one of two groups: group 1 (n = 38) patients had primary infections, whereas group 2 (n = 24) patients had been previously exposed to the microorganism, i.e. re-infection of group 2 individuals occurred after long periods of time during which no active infection by Brucella had been detected. Receivingoperating charts (ROC) were used to determine the diagnostic value of the different tests and to establish discriminant values. Blood culture was a valuable diagnostic tool in group 1 (0.92 sensitivity) but was inappropriate in group 2 (0.08). The combination of positive rose Bengal test and agglutination ≥1/160 was valid for diagnosis in group 1. In group 2, agglutination < 1/160 (including negative agglutination) did not rule out brucellosis. The combination of positive rose Bengal test and Coombs" test ≥1/320 was the best diagnostic criterion (0.8 specificity; 1 sensitivity). ELISA (for IgG, IgM, or both) did not improve diagnostic accuracy

Capsid modified replicating adenovirus to selectively target colon cancer

Résumé: Pratiquement tous les cancers du colon contiennent des mutations dans la voie de signalisation de Wnt qui active constitutivement cette voie. Cette activation mène à la stabilisation de la β-catenine. La β-catenin est transportée dans le noyau ou elle active des gènes cible en interagissant avec le facteur de transcription de TCF/LEF. Des adénovirus qui peuvent sélectivement se répliquer dans les cellules tumorales sont les agents qui peuvent permettre la déstruction de la tumeur mais pas le tissu normal. In vitro, les adénovirus avec des sites d'attachement du facteur de transcription TCF dans les promoteurs de l'adénovirus montrent une sélectivité et une activité dans une large sélection de lignées cellulaires de cancer du colon. Au contraire, in vivo, quand les adénovirus modifiés sont injectés dans la circulation, ils sont moins efficaces à cause de leur fixation par le foie et à cause de l'absence d'expression du récepteur du Coxsackie-Adénovirus (CAR). Le but de ma thèse était de modifier la protéine principale de capside de l'adénovirus, fibre, pour augmenter l'infection des tumeurs du cancer du colon. La fibre de l'adénovirus est responsable de l'attachement aux cellules et de l'entrée virale. J'ai inséré un peptide RGD dans la boucle HI de la fibre qui dirige sélectivement le virus aux récepteurs des integrines. Les integrines sont surexprimées par les cellules du cancer du colon et l'endothélium des vesseaux de la tumeur. Le virus re-ciblé, vKH6, a montré une activité accrue dans toutes les lignées cellulaires de cancer du colon, tandis que la sélectivité était maintenue. In vivo, vKH6 était supérieur au virus avec une capside de type sauvage en retardant la croissance de la tumeur. Le virus s'est répliqué plus vite et dispersé graduellement dans la tumeur. Cet effet a été montré par hybridation in situ et par PCR quantitative. Cependant, la monothérapie avec le virus n'a pu retarder la croissance des cellules tumorales SW620 greffées que de 2 semaines, mais à cause des régions non infectées la tumeur n'a pas pu être éliminée. Bien que la combinaison avec les chimiothérapies conventionnelles soit d'intérêt potentiel, presque toutes interfèrent avec la réplication virale. Les drogues antiangiogéniques sont des agents anti-tumoraux efficaces et prometteurs. Ces drogues n'interfèrent pas avec le cycle de vie de l'adénovirus. RAD001 est un dérivé de la rapamycine et il inhibe mTOR, une protéine kinase de la voie de PI3K. RAD001 empêche la croissance des cellules et il a aussi des effets anti-angiogénique et immunosuppressifs. RAD001 in vitro n'affecte pas l'expression des gènes viraux et la production virale. La combinaison de VKH6 et RAD001 in vivo a un effet additif en retardant la croissance de la tumeur. Des nouveaux peptides plus efficaces dans le ciblage de l'adénovirus sont nécessaires pour augmenter l'infection des tumeurs. J'ai créé un système de recombinaison qui permettra la sélection de nouveaux peptides dans le contexte du génome de l'adénovirus. Summary Virtually all colon cancers have mutations in the Wnt signalling pathway which result in the constitutive activation of the pathway. This activation leads to stabilization of β-catenin. β-catenin enters the nucleus and activates its target genes through interaction with the TCF transcription factor. Selectively replicating adenoviruses are promising novel agents that can destroy the tumour but not the surrounding normal tissue. In vitro, adenoviruses with TCF binding sites in the early viral promoters show selectivity and activity in a broad panel of viruses but in vivo they are less effective due to the lack of expression of the Coxsackie-Adenovirus receptor (CAR). The aim of my thesis was to modify the major capsid protein of the adenovirus, fibre, to increase the infection of colon tumours. Fibre of adenovirus is responsible for the binding to cells and for the viral uptake. I inserted an RGD binding peptide into the HI loop of fibre that selectively targets the virus to integrins that are overexpressed on tumour cells and on tumour endothelium. The retargeted virus, vKH6, showed increased activity in all colon cancer cell lines while selectivity was maintained. In vivo, vKH6 is superior to a matched virus with a wild type capsid in delaying tumour growth. vKH6 replicates and gradually spreads within the tumour as shown by in situ hybridization and Q-PCR. The virus alone can delay the growth of SW620 xenografts by 2 weeks but due to uninfected tumour regions the tumour cannot be cured. Although combination with conventional chemotherapeutics is of potential interest, almost all of them interfere with the viral replication. Growing evidence supports that anti-angiogenic drugs are effective and promising anti-tumour agents. These drugs interfere less with the viral life cycle. RAD001 is a rapamycin derivative and it blocks mTOR, a protein kinase in the PI3K pathway. RAD001 inhibits cell growth and has strong anti-angiogenic and immunosuppressive effects. RAD001 in vitro does not affect viral gene expression and viral burst size. In vivo vKH6 and RAD001 have an additive effect in delaying tumour growth, but tumour growth is still not completely inhibited. To further increase tumour infection new tumour specific targeting peptides are needed. I created an adenovirus display library that will allow the selection of targeting peptides. This system may also facilitate the production of fibre modified viruses.

Treatment of type B periprosthetic femur fractures with curved non-locking plate with eccentric holes: Retrospective study of 43 patients with minimum 1-year follow-up.

INTRODUCTION: Periprosthetic femur fracture (PFF) is a serious complication after total hip arthroplasty that can be treated using different internal fixation devices. However, the outcomes with curved non-locking plates with eccentric holes in this indication have not been reported previously. The objectives of this study were to determine: (1) the union rate; (2) the complication rate; (3) autonomy in a group of patients with a Vancouver type B PFF who were treated with this plate. HYPOTHESIS: Use of this plate results in a high union rate with minimal mechanical complications. MATERIALS AND METHODS: Forty-three patients with a mean age of 79 years±13 (41-98) who had undergone fixation of Vancouver type B PFF with this plate between 2002 and 2007 were included in the study. The time to union and Parker Mobility Score were evaluated. The revision-free survival (all causes) was calculated using Kaplan-Meier analysis. The average follow-up was 42 months±20 (16-90). RESULTS: Union was obtained in all patients in a mean of 2.4 months±0.6 (2-4). One patient had varus malunion of the femur. The Parker Mobility Score decreased from 5.93±1.94 (2-9) to 4.93±1.8 (1-9) (P=0.01). Two patients required a surgical revision: one for an infection after 4.5 years and one for stem loosening. The survival of the femoral stem 5 years after fracture fixation was 83.3%±12.6%. CONCLUSION: Use of a curved plate with eccentric holes for treating type B PFF led to a high union rate and a low number of fixation-related complications. However, PFF remains a serious complication of hip arthroplasty that is accompanied by high morbidity and mortality rates. LEVEL OF EVIDENCE: Retrospective study, level IV.

The UlaG protein familly defines novel structural and functional motifs grafted on an ancient RNase fold

Background: Bacterial populations are highly successful at colonizing new habitats and adapting to changing environmental conditions, partly due to their capacity to evolve novel virulence and metabolic pathways in response to stress conditions and to shuffle them by horizontal gene transfer (HGT). A common theme in the evolution of new functions consists of gene duplication followed by functional divergence. UlaG, a unique manganese-dependent metallo-b-lactamase (MBL) enzyme involved in L-ascorbate metabolism by commensal and symbiotic enterobacteria, provides a model for the study of the emergence of new catalytic activities from the modification of an ancient fold. Furthermore, UlaG is the founding member of the so-called UlaG-like (UlaGL) protein family, a recently established and poorly characterized family comprising divalent (and perhaps trivalent)metal-binding MBLs that catalyze transformations on phosphorylated sugars and nucleotides. Results: Here we combined protein structure-guided and sequence-only molecular phylogenetic analyses to dissect the molecular evolution of UlaG and to study its phylogenomic distribution, its relatedness with present-day UlaGL protein sequences and functional conservation. Phylogenetic analyses indicate that UlaGL sequences are present in Bacteria and Archaea, with bona fide orthologs found mainly in mammalian and plant-associated Gramnegative and Gram-positive bacteria. The incongruence between the UlaGL tree and known species trees indicates exchange by HGT and suggests that the UlaGL-encoding genes provided a growth advantage under changing conditions. Our search for more distantly related protein sequences aided by structural homology has uncovered that UlaGL sequences have a common evolutionary origin with present-day RNA processing and metabolizing MBL enzymes widespread in Bacteria, Archaea, and Eukarya. This observation suggests an ancient origin for the UlaGL family within the broader trunk of the MBL superfamily by duplication, neofunctionalization and fixation. Conclusions: Our results suggest that the forerunner of UlaG was present as an RNA metabolizing enzyme in the last common ancestor, and that the modern descendants of that ancestral gene have a wide phylogenetic distribution and functional roles. We propose that the UlaGL family evolved new metabolic roles among bacterial and possibly archeal phyla in the setting of a close association with metazoans, such as in the mammalian gastrointestinal tract or in animal and plant pathogens, as well as in environmental settings. Accordingly, the major evolutionary forces shaping the UlaGL family include vertical inheritance and lineage-specific duplication and acquisition of novel metabolic functions, followed by HGT and numerous lineage-specific gene loss events.

Ruuvipurkaimen väsymistarkastelu

Ruuvipurkaimen väsymismitoituksen hankalin vaihe on oikeiden kuormien määrittäminen. Tässä työssä laskettiin ruuvipurkaimen väsymisikä kenttämittauksissa saatujen kuormien perusteella. Laskentaan käytettiin nimellisjännitysten menetelmää, Hot-Spot-menetelmää, murtumismekaniikkaa ja paikallisen venymän menetelmää. Purkaimen ruuviputken hitsit käsitellään koneviilaamalla. Kirjallisuudesta etsittiin koetuloksia koneviilatuille kappaleille ja niiden pohjalta laskettiin ehdotus suunnitteluväsymisrajaksi käytettäväksi koneviilatuille T-liitoksille ruuvipurkaimien suunnittelussa. Väsymisrajaksi saatiin 95MPa, kun huomioitiin koekappaleiden koko tilastollisin menetelmin.

Sähkömoottorin rakenteen analysointi tärisevässä käytössä

Tämän työn tavoitteena oli suunnitella esimerkkirakenne vaikeisiin olosuhteisiin tarkoitetusta sähkömoottorista. Ensisijaisesti tarkasteltavia osia sähkömoottorin rakenteessa olivat roottoriakseli, laakerointi sekä laakerikilvet. Laakeroinnista tarkasteltiin laakerivaurioiden syntymistä, tarvittavaa esijännitysvoimaa sekä jälkivoitelua. Lisäksi momenttiakselista tarkasteltiin sen vääntövärähtelyominaisuuksia sekä rungosta ja laakerikilvistä niiden jännityksiä B5-kiinnitysasennossa, eli laippakiinnityksessä. Työstä pyrittiin tekemään suunnitteluohje vastaavanlaisiin suunnittelutehtäviin. Tämän työn kirjallisessa osassa tarkasteltiin oikosulkumoottorin rakennetta ja laakerointia, josta perehdyttiin erityisesti erilaisiin kuormitustilanteisiin, voiteluun sekä kestoiänlaskentaan. Lisäksi tarkasteltiin roottoridynamiikkaa ja ominaismuotojen laskentaa. Kokonaisen roottorin, sisältäen roottoriakselin, –levypaketin, laakeroinnin ja ylikuormitussuojan, pienimmäksi ominaistaajuudeksi saatiin 62,1 Hz:ä, joka oli kuitenkin lähes kaksinkertainen verrattaessa roottorin käyntinopeuteen (33,3 Hz:ä). On siis varmaa, että roottorin kestoikä ei heikkene sen ominaistaajuuksien aiheuttamasta resonanssista. Pelkän roottoriakselin pienimmäksi ominaistaajuudeksi saatiin 448,0 Hz:ä. Verrattaessa tätä taajuutta roottorin käyntinopeuteen nähtiin, että roottoriakselin ominaistaajuuksien puolesta rakenne oli selvästi alikriittinen. Tarkasteltaessa roottoriakselin vääntövärähtelyn ominaistaajuutta 12,6 Hz:ä nähtiin, että se oli pienempi kuin moottorin pyörimisnopeus. Tämä ei kuitenkaan ollut moottorin kestoiän kannalta vakavaa. Koska todellisista sähkömoottorin käyttöolosuhteista ei ollut tarkkaa tietoa, tutkittiin laakeroinnin kestoikää kahdessa ääriolosuhteessa, joko moottoriin kohdistuva tärinä oli kokoajan suurin mahdollinen tunnettu tai tärinää ei ollut ollenkaan. Todellinen tilanne on jossakin näiden kahden ääriolosuhteen välissä. Oli kuitenkin varmaa, että tarkasteltava rakenne kestää halutun kestoiän eli 10000 tuntia laakeroinnin puolesta. Laakerikilpien ominaistaajuudet olivat korkeita eikä niillä ollut vaikutusta rakenteen kestoikään. Laakerikilpien korkeat ominaistaajuudet johtuivat niiden jäykästä rakenteesta ja suuresta materiaalipaksuudesta. Tarkasteltaessa momenttiakselin vääntövärähtelyn ominaistaajuutta 7,2 Hz:ä nähtiin, että se oli pienempi kuin moottorin pyörimisnopeus. Tämä ei kuitenkaan ollut moottorin kestoiän kannalta vakavaa, jos ominaistaajuus ja sen kerrannaiset ohitettaisiin sähkömoottorin käytössä hetkellisesti eikä moottoria käytettäisi vääntövärähtelyn taajuudella. Tarkasteltaessa B5-asennon jännityksiä nähdään, että ne kasvoivat rungon ja laakerikilven välisissä kiinnitysruuveissa yllättävän suuriksi. Normaalin ruuvin murtoraja on 640 MPa:a, kun jännitys rungon ja laakerikilven välisissä kiinnitysruuveissa oli suurimmillaan 400 MPa:a. Rakenne oli kuitenkin varmalla puolella kestävyyden suhteen, koska keskimääräinen vetojännitys ruuveissa oli noin 200 MPa:a. Ruuvien jännityksiä voidaan pienentää lisäämällä kiinnitys-ruuvien lukumäärää tai kasvattamalla niiden kokoa. Koko moottorin kiinnitysruuveissa jännitys oli vain 4 MPa:a, koska osan kuormituksesta kantoi laakerikilven olakkeet.

Rooting and acclimatization of the Japanese plum tree, cv. América

Rooting and acclimatization are limiting steps in plant micropropagation, especially in woody plant species. This study aimed to evaluate the IAA and IBA effect on the in vitro rooting and acclimatization of micropropagated shoots of Japanese plum (Prunus salicina Lindl.) cv. América. Shoots from 3 to 4 cm long were inoculated in MS medium with half salt and vitamin concentrations (MS/2) added with IAA and IBA (0, 0.25, 0.5, 0.75 and 1 mg L-1). After a 20-day period in in vitro cultivation, the shoots were evaluated, and then transferred to a greenhouse, and evaluated after 30 days. At the end of the in vitro cultivation period, no significant interactions were observed for number of roots per shoot and rooting percentage, but a significant effect was recorded for auxin type only, for which shoots grown in media added with IBA showed high values - 0.87 and 41.95%, respectively. A linear increase response from 1.45 to 5.75 cm was verified for root length of shoots cultivated in IBA medium; however, no significant effect was observed, and a 0.86 cm average root length per shoot grown in medium added with IAA was found. After 30 days of acclimatization period, the largest survival percentage was obtained from shoots cultivated in medium with 1 mg L-1 of IBA and IAA (88% and 92%, respectively). Although, IBA provided the highest in vitro rooting, most of the surviving shoots were those originated in IAA-added medium, probably because IBA promoted longer fibrous roots, less appropriate for transplant and soil fixation, as they are easily damaged. It was concluded that in vitro rooting with the addition of the highest IAA concentration (1 mg L-1) provided the greatest plant survival during the acclimatization period of the Japanese plum cv. América.

Soybean (Glycine max. L.) and bacteroid glyoxylate cycle activities during nodular senescence.

Soybean (Glycine max. L.) nodular senescence results in the dismantling of the peribacteroid membrane (PBM) and in an increase of soybean isocitrate lyase (ICL; EC 4.1.3.1) and malate synthase (MS; EC 4.1.3.2) mRNA and protein levels. This suggests that in senescing soybean nodular cells, the specific glyoxylate cycle enzyme activities might be induced to reallocate carbon obtained from the PBM degradation. In order to evaluate as well the carbon metabolism of the nitrogen-fixing Bradyrhizobium japonicum endosymbiotic bacteroids during nodular senescence, their glyoxylate cycle activities were also investigated. To this end, partial DNA sequences were isolated from their icl and ms genes, but the corresponding mRNAs were not detected in the microorganisms. It was also observed that the bacteroid ICL and MS activities were negligible during nodular senescence. This suggests that glyoxylate cycle activities are not reinitiated in the bacteroids under these physiological conditions. In case the microorganisms nevertheless feed on the PBM degradation products, this might occur via the citric acid cycle exclusively.

Effect of biogeographic history on population vulnerability in European amphibians.

The genetic diversity of populations, which contributes greatly to their adaptive potential, is negatively affected by anthropogenic habitat fragmentation and destruction. However, continental-scale losses of genetic diversity also resulted from the population expansions that followed the end of the last glaciation, an element that is rarely considered in a conservation context. We addressed this issue in a meta-analysis in which we compared the spatial patterns of vulnerability of 18 widespread European amphibians in light of phylogeographic histories (glacial refugia and postglacial routes) and anthropogenic disturbances. Conservation statuses significantly worsened with distances from refugia, particularly in the context of industrial agriculture; human population density also had a negative effect. These findings suggest that features associated with the loss of genetic diversity in post-glacial amphibian populations (such as enhanced fixation load or depressed adaptive potential) may increase their susceptibility to current threats (e.g., habitat fragmentation and pesticide use). We propose that the phylogeographic status of populations (i.e., refugial vs. post-glacial) should be considered in conservation assessments for regional and national red lists.

Kerrosleijukattilan polttoaineensyötön ja ilmasäätöjen tutkiminen

Tässä tutkimuksessa käsitellään kerrosleijukattilan kiinteän polttoaineen syöttöjärjestelmän toimintaa ja ilmanjaon vaikutusta kattilan toimintaan. Polttoainejärjestelmän tutkimisessa arvioidaan eri tekijöiden merkitystä höyrynkehityksen huojunnan aiheuttajana. Työllä pyritään tuomaan selvyyttä polttoainejärjestelmän toiminnallisiin peruskysymyksiin kuten turpeen ja kuoren väliseen lajittumiseen, ruuvipurkaukseen ja määrämittauksiin. Työssä esitetään parannusehdotus polttoaineensyötön toiminnan tasoittamiseksi. Ilmasäätöjen osalta on keskitytty tutkimaan erityisesti ilmakanavarakenteiden vaikutusta ilman jakautumiseen. Kattilan ongelmana olivat vasemman ja oikean puolen väliset erot savukaasujen O2-pitoisuuksissa, tulistuslämpötiloissa ja lieriön pinnan korkeudessa. Ilmanjakoon tehtyjen muutosten vaikutuksesta kyseiset erot tasoittuivat huomattavasti. Työssä on lisäksi tarkasteltu mahdollisuuksia kattilan ilmapäästöarvojen parantamiseen erityisesti NOx- ja CO- päästöjen osalta.

EMS mutagenesis in mature seed-derived rice calli as a new method for rapidly obtaining tilling mutant populations.

Background: TILLING (Targeting Induced Local Lesions IN Genomes) is a reverse genetic method that combines chemical mutagenesis with high-throughput genome-wide screening for point mutation detection in genes of interest. However, this mutation discovery approach faces a particular problem which is how to obtain a mutant population with a sufficiently high mutation density. Furthermore, plant mutagenesis protocols require two successive generations (M1, M2) for mutation fixation to occur before the analysis of the genotype can begin. Results: Here, we describe a new TILLING approach for rice based on ethyl methanesulfonate (EMS) mutagenesis of mature seed-derived calli and direct screening of in vitro regenerated plants. A high mutagenesis rate was obtained (i.e. one mutation in every 451 Kb) when plants were screened for two senescence-related genes. Screening was carried out in 2400 individuals from a mutant population of 6912. Seven sense change mutations out of 15 point mutations were identified. Conclusions: This new strategy represents a significant advantage in terms of time-savings (i.e. more than eight months), greenhouse space and work during the generation of mutant plant populations. Furthermore, this effective chemical mutagenesis protocol ensures high mutagenesis rates thereby saving in waste removal costs and the total amount of mutagen needed thanks to the mutagenesis volume reduction.

GENETIC DIVERSITY AND STRUCTURE OF Oenocarpus mapora GERMPLASM CONSERVED AT EASTERN AMAZON

ABSTRACT The aim of this study was to evaluate the genetic diversity and structure in the germoplasm of Oenocarpus mapora conserved at Eastern Amazon. Thus, 88 individuals were genotyped with five microsatellite loci. These individuals belong to 24 accessions that were sampled in eight sample places of three Brazilian Amazon states conserved at the Active Germplasm Bank (AGB) of Embrapa Eastern Amazon. All loci were polymorphic and they generated 85 alleles with an average of 17 alleles per loci. Total genetic diversity (HE) was 0.48. Sample places were considered genetically distinct, with ?p = 0.354. The analysis of molecular variance (AMOVA) identified that the genetic portion among areas was of 36.14% and within 63.86%. The Nei distances varied from 0.091 between Abaetetuba and Santo Antônio do Tauá, both in the state of Pará (PA), to 4.18, between Parintins, AM and Rio Branco, AC. By means of Bayesian analysis, it was identified nine clusters that compose the accessions of the germplasm bank, with different distributions among individuals. The study showed high fixation rates per sample area, which indicates that there may have been significant inbreeding or crossing among parental individuals. It suggests that future samples should be made of different plants in natural populations. Even though, it was verified that there is considerable genetic variation in the germplasm of O. mapora.

An overview of the metabolic differences between Bradyrhizobium japonicum 110 bacteria and differentiated bacteroids from soybean (Glycine max) root nodules: An in vitro 13C-and 31P-nuclear magnetic resonance spectroscopy study

Bradyrhizobium japonicum is a symbiotic nitrogen-fixing soil bacteria that induce root nodules formation in legume soybean (Glycine max.). Using 13C- and 31P-nuclear magnetic resonance (NMR) spectroscopy, we have analysed the metabolite profiles of cultivated B.japonicum cells and bacteroids isolated from soybean nodules. Our results revealed some quantitative and qualitative differences between the metabolite profiles of bacteroids and their vegetative state. This includes in bacteroids a huge accumulation of soluble carbohydrates such as trehalose, glutamate, myo-inositol and homospermidine as well as Pi, nucleotide pools and intermediates of the primary carbon metabolism. Using this novel approach, these data show that most of the compounds detected in bacteroids reflect the metabolic adaptation of rhizobia to the surrounding microenvironment with its host plant cells.

Metabolic and structural rearrangement during dark-induced autophagy in soybean (Glycine max L.) nodules: An electron microscopy and P-31 and C-13 nuclear magnetic resonance study

The effects of dark-induced stress on the evolution of the soluble metabolites present in senescent soybean (Glycine max L.) nodules were analysed in vitro using C-13- and P-31-NMR spectroscopy. Sucrose and trehalose were the predominant soluble storage carbons. During dark-induced stress, a decline in sugars and some key glycolytic metabolites was observed. Whereas 84% of the sucrose disappeared, only one-half of the trehalose was utilised. This decline coincides with the depletion of Gln, Asn, Ala and with an accumulation of ureides, which reflect a huge reduction of the N-2 fixation. Concomitantly, phosphodiesters and compounds like P-choline, a good marker of membrane phospholipids hydrolysis and cell autophagy, accumulated in the nodules. An autophagic process was confirmed by the decrease in cell fatty acid content. In addition, a slight increase in unsaturated fatty acids (oleic and linoleic acids) was observed, probably as a response to peroxidation reactions. Electron microscopy analysis revealed that, despite membranes dismantling, most of the bacteroids seem to be structurally intact. Taken together, our results show that the carbohydrate starvation induced in soybean by dark stress triggers a profound metabolic and structural rearrangement in the infected cells of soybean nodule which is representative of symbiotic cessation.

Lietteenkäsittelyn toimintaan vaikuttavien muutosten hallinta

Object of the Master’s thesis was to obtain clarification to problems related to sludge treatment at the waste water treatment plant at Stora Enso Varkaus Mill. From time to time these problems have caused emissions to exceed given limits. Case was studied by examining existing data, fiber length fractions and experimental methods. Changes at the Mill have reduced total solids emissions. At the same time the requirement for tertiary treatment has grown. Treatment of tertiary sludge is hard. In the future emission limits shall tighten and the urge for tertiary treatment will grow. The significance of thesis’s results may have a great impact to Stora Enso Varkaus Mill in the future. The results give valuable information to follow-up research and guidelines to sludge treatment. The results encourage observing researched matters at longer period of time. Future recommendations emphasize the meaning of maintenance and systematic, appropriate experiments.